In vitro growth and proliferation of multipotent neural stem cells and their progeny

A method for the in vitro proliferation and differentiation of neural stem cells and stem cell progeny comprising the steps of (a) isolating the cells from a mammal, (b) exposing the cells to a culture medium containing a growth factor, (c) inducing the cells to proliferate, and (d) inducing the cells to differentiate is provided.

Skip to:  ·  Claims  ·  References Cited  · Patent History  ·  Patent History

Claims

1. A method for preparing a population of mammalian neural cells enriched with multipotent neural stem cells comprising:

(a) obtaining a population of mammalian neural cells which contains at least one multipotent neural stem cell capable of producing progeny that are capable of differentiating into neurons and glia, including astrocytes;
(b) preparing a culture medium containing one or more predetermined growth factors capable of inducing multipotent neural stem cell proliferation; and
(c) combining the population of mammalian neural cells obtained in (a) with the culture medium prepared in (b) and culturing said mammalian neural cells under conditions that allow the proliferation of said at least one multipotent neural stem cell to produce multipotent neural stem cell progeny which includes daughter multipotent neural stem cells to produce a cell culture that contains a percentage of multipotent neural stem cells that is at least ten fold higher than that of said population of mammalian neural cells obtained in (a).

2. The method of claim additionally comprising:

(d) preparing at least one subsequent cell culture by combining said multipotent neural stem cell progeny produced in (c) with fresh culture medium containing one or more predetermined growth factors which induces multipotent neural stem cell proliferation to proliferate said daughter multipotent neural stem cells to produce a non-primary cell culture that contains a percentage of multipotent neural stem cells that is at least ten fold higher than that of said cell culture prepared in (c).

3. A The method of claim 1 wherein the culture medium prepared in (b) is defined.

4. The method of claim 1 wherein the growth factor in the culture medium prepared in (b) is selected from the group consisting of epidermal growth factor, amphiregulin, fibroblast growth factor and transforming growth factor alpha.

5. The method of claim 1 wherein said population of mammalian neural cells is derived from mammalian neural tissue selected from the group consisting of cerebral cortex tissue, cerebellum tissue, midbrain tissue, brainstem tissue, spinal cord tissue, ventricular tissue and combinations thereof.

6. The method of claim 1 wherein said population of mammalian neural cells obtained in (a) is derived from juvenile or adult neural tissue.

7. The method of claim 1 wherein said population of mammalian neural cells obtained in (a) is derived from human tissue.

8. A method for inducing the in vitro proliferation of a multipotent neural stem cell comprising:

(a) obtaining cells derived from juvenile or adult mammalian neural tissue containing at least one multipotent neural stem cell capable of producing progeny that are capable of differentiating into neurons and glia;
(b) preparing a culture medium containing one or more predetermined growth factors capable of inducing multipotent neural stem cell proliferation; and
(c) preparing a cell culture by combining the cells obtained in (a) with the culture medium prepared in (b) to induce proliferation of said multipotent neural stem cell to produce multipotent neural stem cell progeny which includes daughter multipotent neural stem cells.

9. The method of claim 8 wherein said mammalian neural tissue is ventricular tissue obtained from a mammal that has been administered a growth factor to induce in vivo proliferation multipotent neural stem cells in said ventricular tissue.

10. The method of claim 8 further comprising:

(d) preparing at least one subsequent cell culture by combining said multipotent neural stem cell progeny with fresh culture medium containing one or morepredetermined growth factor which induces multipotent neural stem cell proliferation to proliferate said daughter multipotent neural stem cells to produce more progeny which include more daughter multipotent neural stem cells.

11. The method of 10 wherein step (d) is repeated.

12. The method of claim 8 wherein said growth factor in the culture medium prepared in (b) is selected from the group consisting of epidermal growth factor, amphiregulin, acidic fibroblast growth factor, basic fibroblast growth factor, transforming growth factor alpha, and combinations thereof.

13. The method of claim 8 wherein said growth factor in the culture medium prepared in (b) is selected from the group consisting of epidermal growth factor and transforming growth factor alpha.

14. The method of claim 8 wherein said growth factor in the culture medium prepared in (b) is a fibroblast growth factor.

15. The method of claim 14 wherein said culture medium prepared in (b) additionally contains epidermal growth factor.

16. The method of claim 8 wherein said mammalian neural tissue is selected from the group consisting of cerebral cortex tissue, cerebellum tissue, midbrain tissue, brainstem tissue, spinal cord tissue and ventricular tissue.

17. The method of claim 8 wherein said mammalian neural tissue is obtained from a human.

18. The method of claim 8 wherein said culture medium is substantially serum-free and wherein the cells obtained in (a) have not been cultured in a serum-containing medium.

19. The method of claim 8 wherein said culture medium is defined.

20. The method of claim 8 further comprising:

(d) inducing the multipotent neural stem cell progeny to differentiate to produce a cell culture comprising differentiated neural cells.

21. A method for inducing the in vitro proliferation of a multipotent neural stem cell comprising:

(a) obtaining cells derived from mammalian neural tissue containing at least one multipotent neural stem cell capable of producing progeny that are capable of differentiating into neurons and glia, including astrocytes, wherein said obtained cells have not been cultured in a serum-containing medium;
(b) preparing a substantially serum-free culture medium containing at one or more predetermined growth factors capable of inducing proliferation of said multipotent neural stem cell;
(c) preparing a cell culture by combining the cells obtained in (a) with the culture medium prepared in (b) to induce proliferation of said multipotent neural stem cell to produce multipotent neural stem cell progeny which includes daughter multipotent neural stem cells; and
(d) preparing at least one subsequent cell culture by combining said multipotent neural stem cell progeny with fresh substantially serum-free culture medium containing at least one or more predetermined growth factors which induces multipotent neural stem cell proliferation to proliferate said daughter multipotent neural stem cells to produce more progeny which include more daughter multipotent neural stem cells.

22. The method of claim 21 wherein growth factor in the culture medium prepared in (b) is selected from the group consisting of epidermal growth factor, amphiregulin, acidic fibroblast growth factor, basic fibroblast growth factor, transforming growth factor alpha, and combinations thereof.

23. The method of claim 21 wherein said mammalian neural tissue is ventricular tissue obtained from a mammal that has been administered a growth factor to induce in vivo proliferation of said multipotent neural stem cell in said ventricular tissue to produce multipotent neural stem cell progeny.

24. The method of claim 21 wherein the source of said neural tissue is a transgenic animal.

25. The method of claim 21 wherein said mammalian neural tissue is from a mouse.

26. The method of claim 21 wherein said mammalian neural tissue is from a primate.

27. The method of claim 21 wherein the source of said neural tissue is a human.

28. The method of claim 21 wherein said mammalian neural tissue is derived from a juvenile or adult mammal.

29. The method of claim 21 wherein said mammalian neural tissue is selected from the group consisting of cerebral cortex tissue, cerebellum tissue, midbrain tissue, brainstem tissue, spinal cord tissue and ventricular tissue.

30. The method of claim 21 wherein said growth factor in the culture medium prepared in (b) is selected from the group consisting of epidermal growth factor and transforming growth factor alpha.

31. The method of claim 21 wherein said growth factor in the culture medium prepared in (b) is a fibroblast growth factor.

32. The method of claim 31 wherein said culture medium prepared in (b) additionally contains epidermal growth factor.

33. The method of claim 21 wherein the culture media prepared in (b) and (d) are defined.

34. The method of claim 21 wherein said culture medium prepared in (b) additionally contains a regulatory factor that regulates the proliferation of said multipotent neural stem cell proliferated in (c) and/or regulates proliferation of said multipotent neural stem cell progeny.

35. The method of claim 34 wherein said regulatory factor is different from said growth factor in the culture medium prepared in (b) and is selected from the group consisting of heparan sulfate, a member of the transforming growth factor beta family, ciliary neurotrophic factor, retinoic acid, activin, interleukins, the Bcl-2 gene product, platelet-derived growth factor, nerve growth factor, a macrophage inflammatory protein, tumor necrosis factor alpha and epidermal growth factor.

36. The method of claim 21 wherein said multipotent neural stem cell progeny produced in (c) and (d) are in suspension.

37. The method of claim 36 wherein in (c) the progeny of a single multipotent neural stem forms a clonally-derived cluster of cells.

38. The method of claim 37 wherein prior to (d) said clonally-derived cluster of cells is dissociated to form a suspension comprising single cells which are transferred to said fresh culture medium in (d).

39. The method of claim 21 further comprising:

(e) inducing the muitipotent neural stem cell progeny produced in (d) to differentiate to produce a cell culture comprising differentiated neural cells.

40. The method of claim 39 wherein said multipotent neural stem cell progeny in (e) are induced to differentiate in a culture medium containing at least one growth factor which influences differentiation of said multipotent neural stem cell progeny.

41. The method of claim 5 wherein said growth factor which influences differentiation of said multipotent neural stem cell progeny is selected from the group consisting of ciliary neurotrophic factor, basic fibroblast growth factor, and retinoic acid.

42. The method of claim 39 wherein said multipotent neural stem cell progeny in (e) are contacted with a substrate which induces differentiation of said multipotent neural stem cell progeny.

43. The method of claim 39 wherein said multipotent neural stem cell progeny in (e) are induced to differentiate in suspension by not reinitiating induction of proliferation.

44. The method of claim 39 wherein said multipotent neural stem cell progeny in (e) are induced to differentiate in a culture medium that is substantially free of said growth factor(s) contained in the culture media of (b) and (d).

45. The method of claim 4 wherein said multipotent neural stem cell progeny in (e) are induced to differentiate in a culture medium containing serum.

46. The method of claim 45 wherein said growth factor in the culture medium prepared in (b) is a fibroblast growth factor and said regulatory factor is selected from the group consisting of heparan sulfate, epidermal growth factor, and a combination thereof.

47. A method for the in vitro proliferation of a human multipotent neural stem cell comprising:

(a) obtaining cells derived from human neural tissue containing at least one multipotent neural stem cell capable of producing progeny that are capable of differentiating into neurons and glia, including astrocytes;
(b) preparing a culture medium containing one or more predetermined growth factors capable of inducing multipotent neural stem cell proliferation; and
(c) preparing a cell culture by combining the cells obtained in (a) with the culture medium prepared in (b) to induce proliferation of said multipotent neural stem cell to produce multipotent neural stem cell progeny which includes daughter multipotent neural stem cells.

48. The method of claim 47 wherein cells obtained in (a) have not been cultured in a serum-containing medium, and wherein the culture medium prepared in (b) is substantially serum-free.

49. The method of claim 47 wherein said culture medium is defined.

50. The method of claim 47 wherein said growth factor is selected from the group consisting of epidermal growth factor, amphiregulin, fibroblast growth factor, transforming growth factor alpha, and combinations thereof.

51. The method of claim 47 wherein said human neural tissue is selected from the group consisting of cerebral cortex tissue, cerebellum tissue, midbrain tissue, brainstem tissue, spinal cord tissue and ventricular tissue.

52. The method of claim 47 wherein said growth factor in the culture medium of (b) is a fibroblast growth factor.

53. The method of claim 52 wherein the culture medium of (b) additionally contains epidermal growth factor.

54. The method of claim 47 further comprising:

(d) preparing at least one subsequent cell culture by combining said multipotent neural stem cell progeny with fresh culture medium containing one or more predetermined growth factors which induces multipotent neural stem cell proliferation to proliferate said daughter multipotent neural stem cells to produce more progeny which include more daughter multipotent neural stem cells.

55. A composition comprising a population of non-primary neural cells which are derived from a primary cell culture, said population of non-primary. neural cells having a greater percentage of multipotent neural stem cells compared to that of said primary cell culture, wherein a single multipotent neural stem cell is capable of producing progeny that are capable of differentiating into neurons and glia, including astrocytes.

56. The composition of claim 55 wherein said percentage of multipotent neural stem cells of said population of non-primary neural cells is at least ten fold higher than that of said primary cell culture.

57. The composition of claim 55 which additionally comprises a growth factor selected from the group consisting of epidermal growth factor, amphiregulin, acidic fibroblast growth factor, basic fibroblast growth factor, transforming growth factor alpha, and combinations thereof.

58. The composition of claim 55 which additionally comprises a defined culture medium.

59. The composition of claim 55 wherein said multipotent neural stem cells are derived from juvenile or adult neural tissue.

60. The composition of claim 55 wherein said multipotent neural stem cells are derived from neural tissue selected from the group consisting of cerebral cortex tissue, cerebellum tissue, midbrain tissue, brainstem tissue, spinal cord tissue, ventricular tissue, and combinations thereof.

61. The composition of claim 55 wherein said multipotent neural stem cells are derived from human neural tissue.

62. A composition comprising daughter multipotent neural stem cells produced by the method of claim 8.

63. The composition of claim 62 which additionally comprises a growth factor selected from the group consisting of epidermal growth factor, amphiregulin, acidic fibroblast growth factor, basic fibroblast growth factor, transforming growth factor alpha, and combinations thereof.

64. The composition of claim 62 which additionally comprises a defined culture medium.

65. The composition of claim 62 wherein said daughter multipotent neural stem cells are derived from human neural tissue.

66. The composition of claim 62 wherein said daughter multipotent neural stem cells are derived from neural tissue selected from the group consisting of cerebral cortex tissue, cerebellum tissue, midbrain tissue, brainstem tissue, spinal cord tissue, ventricular tissue, and combinations thereof.

67. A composition comprising multipotent neural stem cell progeny including daughter multipotent neural stem cells produced by the method of claim 21.

68. The composition of claim 67 which additionally comprises a growth factor selected from the group consisting of epidermal growth factor, amphiregulin, acidic fibroblast growth factor, basic fibroblast growth factor, transforming growth factor alpha, and combinations thereof.

69. A composition according to claim 67 which additionally comprises a defined culture medium.

70. The composition of claim 67 wherein said daughter multipotent neural stem cells are derived from neural tissue selected from the group consisting of cerebral cortex tissue, cerebellum tissue, midbrain tissue, brainstem tissue, spinal cord tissue, ventricular tissue, and combinations thereof.

71. The composition of claim 67 wherein said daughter multipotent neural stem cells are derived from juvenile or adult mammalian neural tissue.

72. The composition of claim 67 wherein said daughter multipotent neural stem cells are derived from human neural tissue.

73. A composition comprising differentiated neural cells produced by the method of claim 39.

74. The composition of claim 73 wherein said differentiated neural cells are selected from the group consisting of neurons, type I astrocytes, type II astrocytes, oligodendrocytes, and combinations thereof.

75. A composition comprising daughter multipotent neural stem cells produced by the method of claim 47.

76. The composition of claim 75 which additionally comprises a growth factor selected from the group consisting of epidermal growth factor, amphiregulin, acidic fibroblast growth factor, basic fibroblast growth factor, transforming growth factor alpha, and combinations thereof.

77. The composition of claim 75 which additionally comprises a defined culture medium.

78. The composition of claim 75 wherein said daughter multipotent neural stem cells are derived from juvenile or adult neural tissue.

79. The composition of claim 75 wherein said daughter multipotent neural stem cells are derived from neural tissue selected from the group consisting of cerebral cortex tissue, cerebellum tissue, midbrain tissue, brainstem tissue, spinal cord tissue, ventricular tissue, and combinations thereof.

80. A cluster of cells prepared in vitro consisting of the progeny of a single multipotent neural stem cell, wherein said progeny are capable of differentiating into neurons and glia, including astrocytes.

Referenced Cited
U.S. Patent Documents
4753635 June 28, 1988 Sagen et al.
4980174 December 25, 1990 Sagen et al.
5082670 January 21, 1992 Gage
5175103 December 29, 1992 Lee et al.
5411883 May 2, 1995 Boss et al.
5612211 March 18, 1997 Wilson et al.
Foreign Patent Documents
0 233 838 August 1987 EPX
89/03872 May 1989 WOX
90/06757 June 1990 WOX
91/02003 February 1991 WOX
91/09936 July 1991 WOX
91/17242 November 1991 WOX
93/01275 January 1993 WOX
93/09802 May 1993 WOX
94/03199 February 1994 WOX
Other references
  • Almazan et al., "Epidermal Growth and Bovine Growth Hormone Stimulate Differentiation and Myelination of Brain Cell Aggregates in Culture," Developmental Brain Research, 21:257-264 (1985). Anchan et al., "Trophic Factors Influence Proliferation of Germinal Neuroepithelial Cells of the Retina, " J. Cell Biol., 109:58a, Abstract No. 308 (1989). Anchan et al., "EGF and TGF-.alpha. Stimulate Retinal Neuroepithelial Cell Proliferation in Vitro," Neuron, 6(6):923-936 (1991). Bayer et al., "Neuron production in the Hippocampus and olfactory bulb of the adult rat Brain: addition or replacement?", Annals NY. Acad. Sci. 457:163-172 (1985). Bjorklund et al., "Neural Grafting in Animal Models of Neurodegenerative Diseases," Ann. New York Acad. Sci., 457:53-81 (1985). Bouvier et al., "Basic Fibroblast Growth Factor (bFGF) Promotes the Survival and Proliferation of Mesencephalic Neuronal Precursors in Vitro," Society for Neuroscience Abstracts, vol. 18, Abstract No.: 403.7 (1992). Boyles et al., "Accumulation of Apolipoproteins in the Regenerating and Remyelinating Mammalian Peripheral Nerve," J. Biol. Chem., 265(29):17805-17815 (1990). Calof et al., "Analysis of Neurogenesis in a Mammalian Neuroepithelium: Proliferation and Differentiation of an Olfactory Neuron Precursor in Vitro," Neuron, 3:115-127 (1989). Cattaneo et al., "Identifying and Manipulating neuronal stem cells," TINS, 14(8): 338-340 (1991). Cattaneo et al., "Proliferation and differentiation of neuronal stem cells regulated by nerve growth factor," Nature, 347:762-765 (1990). Cepko "Immortalization of neural cells via retrovirus-mediated oncogene transduction," Ann. Rev. Neurosci., 12:47-65 (1989). Deloulme et al., "Establishment of Pure Neuronal Cultures From Fetal Rat Spinal Cord and Proliferation of the Neuronal Precursor Cells in the Presence of Fibroblast Growth Factor," Journal of Neuroscience Research, 29:499-509 (1991). Dunnett et al., "Dopamine-rich transplants in experimental Parkinsonism," TINS, 266-270 (Jul. 1983). Emerich et al., "Behavioral Effects of Neural Transplantation,"Cell Transplantation, 1:1-27 (1992). Faaland et al., "Rapid uptake of tyrphostin into A431 human epidermoid cells is followed by delayed inhibition of epidermal growth factor (EGF)-stimulated EGF receptor tyrosine kinase activity", Mol. Cell Biol. 11(5):2697-2703 (1991). Ferrari et al., "Basic Fibroblast Growth Factor Promotes the Survival and Development of Mesencephalic Neurons in Culture," Developmental Biology, 133:140-147 (1989). Frederickson et al., "Immortal Neuroepithelial Precursor Cell Lines," Society for Neuroscience Abstracts, 13:182 Abstract No. 55.6 (1987). Frederiksen et al., "Proliferation and differentiation of rat neuroepithelial precursor cells in vivo," The Journal of Neuroscience 8(4):1144-1151 (1988). Frederiksen et al., "Immortalization of precursor cells from the mammalian CNS," Neuron, 1:439-448 (1988). Freed et al., "Transplantation of human fetal dopamine cells for Parkinson's disease," Arch. Neurol., 47:505-512 (1990). Freshney, "Culture of Animal cells--A Manual of Basic Techniques", Alan R. Liss, Inc., N.Y. pp. 190-195. Freshney, "Culture of Animal Cells--A Manual of Basic Techniques," Alan R. Liss, Inc., N.Y., Chapter 11, pp. 137-153 (1987). Geller et al., "A defective HSV-1 vector expresses Escherichia coli .beta.-galactosidase in cultured peripheral neurons," Science, 241:1667-1669 (1988). Gensburger et al., "Brain basic fibroblast growth factor stimulates the proliferation of rat neuronal precursor cells in vitro," FEBS Letts, 217(1):1-5 (1987). Godfraind et al., "In Vivo Analysis of Glial Cell Phenotypes during a Viral Demyelinating Disease in Mice," Journal of Cell Biology 109(5):2405-2416. Groves et al., "Repair of demyelinated lesions by transplantation of purified O-2A progenitor cells," Nature, 362:453 (1993). Hall et al., "Stem cells: the generation and maintenance of cellular diversity," Development, 106:619-633 (1989). Hoffman et al., "Transplantation of a polymer-encapsulated cell line genetically engineered to release NGF," Exp. Neurol. 122:100-106 (1993). Hunter et al., "Growth factor responses of enriched bipotential glial progenitors," Developmental Brain Research, 54(2):235-248 (1990). Hunter et al., "Growth factor responses of enriched bipotential glial progenitors," Biol. Abstr. 90:7, Abstract No. 78581 (1990). Hurtig et al., "Postmortem analysis of adrenal-medulla-to-caudate autograft in a patient with Parkinson's disease," Annals of Neurology, 25(6):607-614 (1989). Jiao et al., "Intracerebral transplants of primary muscle cells: a potential `platform` for transgene expression in the brain," Brain Research, 575:143 (1992). Kaplan, "Neurogenesis in the 3-month-old rat visual cortex," J. Comp. Neurol., 195:323 (1981). Kawaja et al., "Somatic gene transfer of nerve growth factor promotes the survival of axotomized septal neurons and the regeneration of their axons in adult rats," J. Neurosci., 12(7)2849 (1992). Korr et al., "Autoradiographic investigation of glial proliferation in the bran of adult mice," J. Comp. Neurol., 150(2):169-176 (1973). Lendahl et al., "CNS stem cells express a new class of intermediate filament protein," Cell, 60 585-595 (1990). Lin et al., "GDNF: A Glial Cell Line-Derived Neurotrophic Factor Midbrain Dopaminergic Neurons," Science, 260:1130 (1993). Lindvall et al., "Grafts of fetal dopamine dnurons survive and improve motor function in Parkinson's disease," Science, 247:574-577 (1990). Lois et al., "Migration of neuroblasts from the lateral ventricle to the olfactory bulb in the adult mammalian CNS", Society for Neuroscience Abstracts, vol. 19, Abstract #361.6 (1993). Luskin et al., Rate and pattern of migration of olfactory bulb interneurons generated postnatally in the subventricular zone, Society for Neuroscience Abstracts, vol. 19, Abstract #361.9 (1993). Masters et al., "Insulin-like growth factor I (IGF-I) receptors and IGF-I action in oligodendrocytes from rat brains," Regulatory Peptides, 33(2):117-131 (1991). Masters et al., "Insulin-like growth factor I (IGF-I) receptors and IGF-I action in oligodendrocytes from rat brains," Biol. Abstr. 93:3, Abstract No. 31828 (1992). McKinnon et al., "FGF modulates the PDGF-driven pathway of oligodendrocyte development," Neuron 5:603-614 (1990). Morrison et al., "Trophic stimulation of cultured neurons from neonatal rat brain by epidermal growth factor," Science, 238:72-75 (1987). Metcalf, "The hemopoietic regulators--an embarrassmetn of riches", Bioassays 14(12):799-805 (1992). Morshead et al., "Neural stem cells are located in the subependymal region of the adult mammalian forebrain", Society for Neuroscience Abstracts, vol. 19, Abstract#360.7 (1993). Morshead et al., "Postmitotic Death is the Fate of Constitutively Proliferating Cells in the Subependymal Layer of the Adult Mouse Brain," The Journal of Neuroscience, 12(1):249-256 (1992). Murphy et al., "Fibroblast growth factor stimulates the Proliferation and Differentiation of Neural Precursor Cells In Vitro," Jour. of Neuroscience Research, 25(4):463-475 (1990). Mytilineou et al., "Epidermal Growth Factor-Induced Survival and Proliferation of Neuronal Precursor Cells from Embryonic Rat Mesencephalon," Neuroscience Letters, 135:62-66 (1992). Nakafuku et al., "Epidermal growth factor and transforming growth factor-.alpha. can induce neuronal differentiation of rat pheochromocytoma PC12 cells under particular culture conditions" FEBS Letts, 315(3):227-232 (1993). Notter et al., "Neuronal properties of monkey adrenal medulla in vitro" Cell Tissue Res., 244:69-76 (1986). Pallage et al., "Long-term effects of nerve growth factor and neural transplants on behavior of rats with medial septal lesions," Brain Research, 386:197-208 (1986). Palmer et al., "Genetically modified skin fibroblasts persist long after transplantation but gradually inactivate introduced genes" Proc. Nat'l. Acad. Sci. USA, 88:1330-1334 (1991). Perlow et al., "Brain grafts reduce motor abnormalities produced by destruction of nigrostriatal dopamine system" Science, 204:643-646 (May, 1979). Piszckiewicz et al., "Proliferation and Survival of Rat Sensory Neuron Precursosrs in Vitro," Society for Neuroscience Abstracts, 19:1709 Abstract No. 704.7 (1993). Potten et al., "Stem cells: attributes, cycles, spirals, pitfalls and uncertainties. Lessons for and from the Crypt," Development, 110:1001-1020 (1990). Raff et al., "A glial progenitor cell that develops in vitro into an astrocyte or an oligodendrocyte depending on culture medium", Nature 303:390-396 (1983). Rakic, "Limits of neurogenesis in primates," Science 227:1054 (1985). Ramatowski et al., "Laminin enhances proliferation and migration of EGRF-generated CNS stem cell progeny," Society for Neuroscience Abstracts, vol. 19, Abstract #360.10 (1993). Reh et al., "Age of differentiation determines rat retinal germinal cell phenotype: Induction of differentiation by dissociation," The Journal of Neuroscience, 9 (12): 4179-4189 (1989). Renfranz et al., "Region-specific differentiation of the hippocampal stem cell line HiB5 upon implantation into the developing mammalian brain" Cell 66:713-729 (1991). Reynolds et al., "EGF-responsive stem cells in the mammalian central nervous system," Neuronal Cell Death and Repair Ch. 19, pp. 247 ed. Cuello (1993). Reynolds et al., "A non-transformed, growth factor-dependent stem cell line derived from the embryonic mouse CNS produces neurons, astrocytes and oligodendrocytes," Restrorative Neurology and Neuroscience, 4 (3) Abstract No. 34.P3 (1992). Reynolds et al., "A multipotent EGF-responsive striatal embryonic progenitor cell produces neurons and astrocytes," J. Neurosci. 12(11):4565-4574 (1992). Reynolds et al., "EGF-and TGF.alpha.-responsive striatal embryonic progenitor cells product both neurons and astrocytes" Soc. Neurosc. Abstracts, 16 Abstract No. 474.2 (Oct./Nov. 1990). Reynolds et al., "Generation of neurons and astrocytes form isolated cells of the adult mammalian central nervous system," Science, 255:1707-1710 (1992). Rohrer et al., "Proliferation and Differentiation of Neuronal Precursor Cells in the Chick Peripheral Nervous System," Biol. Chem. Hoppe Seyles, 368(10):1290-1296 (1987). Ronnett et al., "Human cortical neuronal cell line: establishment from a patient with unilateral megalencephaly," Science, 248:603-605 (1990). Rosenberg et al., "Grafting genetically modified cells to the damaged brain: restorative effects of NGF expression," Science, 242:1575-1578 (1988). Sensenbrenner et al., "Proliferation of Neuronal Precursor Cells from the Central Nervous System in Culture," Reviews in the Neurosciences, 5:43-53 (1994). Smart, "The subependymal layer of the mouse brain and its cell production as shown by radioautography after Thymidine-H.sup.3 injection", J. Comp. Neurol. 116:325 (1961). Snyder et al., "Multipotent neural cell lines can engraft and participate in development of mouse cerebellum", Cell 68:33-51 (1992). Soreto et al., in Neurochemistry: A Practical Approach, chapter 2, pp. 27-63 (1987). Steinbusch et al., "Basic fibroblast growth factor enhances survival and sprouting of fetal dopaminergic cells implanted in the denervated rat caudate-putamen: preliminary observations," Progress in Brain Research, 82:81-86 (1990). Temple "Division and differentiation of isolated CNS blast cells in microculture," Nature, 340:471-473 (1989). Travis, "The search for liver stem cells picks up", Science 259:1829 (1993). Van Der Maazen, et al., "Radiosensitivity of Glial Progenitor Cells of the Perinatal and Adult Rat Optic Nerve Studied by and In-vitro Clonogenic Assay", Biosis Abstract No. 91:324328, Radiother. Oncol 20. (1991). Vescovi et al., "Continual proliferation of EGF-dependent progenitor cells of the embryonic human CNS in vitro", Society for Neuroscience Abstracts, vol. 19, Abstract #360.12 (1993). Walsh et al., "Clonally related cortical cells show several migration patterns", Science, 241:1342 (1988). Watts et al., "Adrenal-caudate transplantation in patients with Parkinson's disease (PD):--1-year follow-up," Neurology 39 (Suppl. 1) Abstract No. PP72 (1989). Weiss et al., "Synaptogenesis of cultured striatal neurons in serum-free medium: a morphological and biochemical study," Proc. Natl. Acad. Sci. USA, 83:2238-2242 (1986). Williams "Programmed Cell Death: Apoptosis and Oncogenesis," Cell, 1097 (1991). Williams et al., "Continuous infusion of nerve growth factor prevents basal forebrain neuronal death after fimbria fornix transection", P.N.A.S. 83:9231 (1986). Widner et al., "Bilateral fetal mesencephalic grafting in two patients with parkinsonism induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)", New Eng. J. Med. 327(22):1556 (1992). Wolff et al., "Grafting Fibroblasts Genetically Modified to Produce L-dopa in a Rat Model of Parkinson Disease," PNAS, 86:9011-9014 (1989). Wolswijk et al., "Identification of an adult-specific glial progenitor cell" Development, 105:387-400 (1989). Yamada et al., Growth of cells in hormonally defined media--Book A Cold Spring Harbor Laboratory, 1982--Cold Spring Harbor conferences on cell proliferation, vol. 9, 131-143. Nurcombe et al., "Developmental Regulation of Neural Response to FGF-1 and FGF-2 By Heparan Sulfate Proteoglycan," Science, 260:103-106 (1993). Brickman et al., "Heparan Sulfates Mediate the Binding of Basic Fibroblast Growth Factor to a Specific Receptor on Neural Precursor Cells," Journal of Biological Chemistry, 270(42):24941-24948 (1995). Blakemore et al., "Extensive Oligodendrocyte Remyelination Following Injection of Cultured Central Nervous System Cells into Demyelinating Lesions in Adult Central Nervous System," Developmental Neuroscience, 10:1-11 (1988). Pezzoli et al., "Intraventricular Infusion of Epidermal Growth Factor Restores Dopaminergic Pathway in Hemiparkinsonian Rats," Movement Disorders, 6(4):281-287 (1991). Stenevi et al., "Effects of Localized Intracerebral Injections of Nerve Growth Factor on the Regenerative Growth of Lesioned Central Noradrenergic Neurones," Brain Research, 69(2):217-234 (1974). Palella et al., "Expression of Human HRPT mRNA in Brains of Mice Infected with a Recombinant Herpes Simplex Virus-1 Vector," Gene, 80:137-144 (1989). Olson, "Grafts and Growth Factors in CNS: Basic Science with Clinical Promise," Proceedings of the Xth Meeting of the World Society for Stereotactic and Functional Neurosurgery, Macbashi: Japan. October 1989, Stereotact Funct. Neurosurg., 54-55:250-167 (1990). Jackowski, "Neural Injury Repair: Hope for the Future as Barriers to Effective CNS Regeneration Become Clearer," British Journal of Neurosurgery, 9:303-317 (1995). Lubetzki et al., "Gene Transfer of Rat Mature Oligodendrocytes and Glial Progenitor Cells with the LacZ Gene," Ann. New York Acad. Sci., 605:66-70 (1990). Friedmann, "Gene Therapy for Neurological Disorders," TIG, 10(6):210-214 (1994). Orkin et al., "Report and Recommendations of the Panel to Assess the NIH Investment in Research on Gene," NIH, pp. 1-40 (Dec. 7, 1995). Kumar et al., "Identification of a Set of Genes with Developmental Down-Regulated Expression in the Mouse Brain," Biochemical and Biophysical Research Comm., 185(3):1155-1161 (1992). Stratagene, 1991 Product Catalog, pp. 115-116. Sambrook et al., "Molecular Cloning: A Laboratory Manual," 2nd ed., Cold Spring Harbor Press 12.2-12.10 (1989). Zecchinelli et al., "Epidermal Growth Factor (EGF) Enhances, In Rats, Dopaminergic Pathway In-vivo an Immunohistochemical Study," Socy, Neurosciences Abstracts, abstract 413.17 (Nov. 1, 1990). Lo et al., "V-myc Immortalizationof Early Rat Neural Crest Cells Yields a Clonal Cell Line Which Generates Both Glial and Adrenergic Progenitor Cells," Developmental Biology, 145:139-153 (1991). Anchan et al., Neuron, 6: 923-936, 1991. Lin et al., Science, 260: 1130-1132, 1993. Ferrari et al., Dev. Biol., 133: 140-147, 1989. Morshead et al., J. Neuroscience, 12(1):249-256, 1992. Hunter et al., Dev. Brain Res., 43: 235-248, 1990.
Patent History
Patent number: 5851832
Type: Grant
Filed: Jun 7, 1995
Date of Patent: Dec 22, 1998
Assignee: Neurospheres, Ltd.
Inventors: Samuel Weiss (Alberta), Brent Reynolds (Alberta), Joseph P. Hammang (Barrington, RI), E. Edward Baetge (Barrington, RI)
Primary Examiner: George C. Elliott
Assistant Examiner: Johnny F. Railey, II
Law Firm: Flehr Hohbach Test Albrition & Herbert LLP
Application Number: 8/486,648